Haifei He

Down-regulation of microRNA-200c is associated with drug resistance in human breast cancer

Drug resistance remains a major clinical obstacle to successful treatment in breast cancer patients, and the evidence of microRNAs involvement in cancer drug resistance has been emerging recently. However, the role of microRNA-200c (miR-200c) in modulating chemoresistance of breast cancer remains largely unexplored. Here, we investigated the miR-200c expression in tumor specimens obtained from thirty-nine breast cancer patients who received neoadjuvent chemotherapy by quantitative real-time PCR. Down-regulated miR-200c was observed in non-responders as compared to responders. In addition, miR-200c expression was observed to be down-regulated over 800-fold in human breast cancer cells resistant to doxorubicin MCF-7/ADR as compared to the parental MCF-7 cells. Up-regulation of miR-200c with transfection of miR-200c mimics in breast cancer cells could enhance the chemosensitivity to epirubicin and reduce expression of multidrug resistance 1 mRNA and P-glycoprotein. Moreover, our study demonstrated that restoration of miR-200c in MCF-7/ADR cells could increase intracellular doxorubicin accumulation determined by flow cytometry. Taken together, our findings suggest that miR-200c may act as a promising therapeutic target for improvement of responsiveness to chemotherapy in breast cancer.

MicroRNAs and drug resistance of breast cancer: basic evidence and clinical applications.

Breast cancer is the most common malignancy and leading cause of cancer mortality in women. Drug resistance is a major obstacle in systemic therapy of breast cancer, which leads therapeutic failure, incontrollable disease, and mortality. MiRNAs are an emerging field in cancer research. Recent evidence demonstrates that miRNAs are core regulators in drug resistance of breast cancer. Preclinical research reveals that miRNAs modulate the multidrug-resistant signal transduction network through up-regulated drug efflux transporters and anti-apoptotic proteins, acquisition of epithelial-mesenchymal transition, and formation of cancer stem cells. MiRNAs mediate endocrine resistance through modulating ERα expression, receptor tyrosine kinase signaling, cell survival signaling, and apoptosis. Such emerging evidence indicates that miRNAs could be potential biomarkers for predicting a response to systemic therapy and prognosis in clinical settings. Targeting specific miRNAs of the drug-resistant network is promising in overcoming drug resistance in breast cancer.

Colorectal cancer lymph node staining by activated carbon nanoparticles suspension in vivo or methylene blue in vitro

AIM To investigate whether activated carbon nanoparticles suspension (ACNS) or methylene blue (MB) can increase the detected number of lymph nodes in colorectal cancer. METHODS Sixty-seven of 72 colorectal cancer patients treated at our hospital fulfilled the inclusion criteria of the study which was conducted from December 2010 to February 2012. Seven patients refused to participate. Eventually, 60 patients were included, and randomly assigned to three groups (20 in each group): ACNS group (group A), MB group (group B) and non-stained conventional surgical group (group C). In group A, patients received subserosal injection of 1 mL ACNS in a 4-quadrant region around the mass. In group B, the main artery of specimen was identified and isolated after the specimen was removed, and 2 mL MB was slowly injected into the isolated, stretched and fixed vessel. In group C, no ACNS and MB were injected. All the mesentery lymph nodes were isolated and removed systematically by visually inspecting and palpating the adipose tissue. RESULTS No difference was observed among the three groups in age, gender, tumor location, tumor diameter, T-stage, degree of differentiation, postoperative complications and peritoneal drainage retention time. The total number of detected lymph nodes was 535, 476 and 223 in the three groups, respectively. The mean number of detected lymph nodes per patient was significantly higher in group A than in group C (26.8 ± 8.4 vs 12.2 ± 3.2, P < 0.001). Similarly, there were significantly more lymph nodes detected in group B than in group C (23.8 ± 6.9 vs 12.2 ± 3.2, P < 0.001). However, there was no significant difference between group A and group B. There were 50, 46 and 32 metastatic lymph nodes dissected in 13 patients of group A, 10 patients of group B and 11 patients of group C, without significant differences among the three groups. Eleven of the 60 patients had insufficient number of detected lymph nodes (< 12). Only one patient with T(4a) rectal cancer had 10 lymph nodes detected in group B, the other 10 patients were all from group C. Based on the different diameter categories, the number of detected lymph nodes in groups A and B was significantly higher than in group C. However, there was no statistically significant difference between group A and group B. The metastatic lymph nodes were not significant different among the three groups. Similarly, tumor location, T stage and tumor differentiation did not affect the staining results. Body mass index was a minor influencing factor in the two different staining methods. The stained lymph nodes can easily be identified from the mesenteric adipose tissues, and the staining time for lymph nodes was not significantly different compared with unstained group. None of the patients in groups A and B had drug-related complications. CONCLUSION Both activated carbon nanoparticles suspension in vivo and methylene blue in vitro can be used as tracers to increase the detected number of lymph nodes in colorectal cancer.

Effect of Age on Breast Cancer Patient Prognoses: A Population-Based Study Using the SEER 18 Database.

Background Age is an important risk factor for breast cancer, but data regarding whether patient age at diagnosis is related to breast cancer survival are conflicting. This population-based study evaluated the effect of age on breast cancer prognosis and identified outcome-related factors. Patients and Methods We searched the Surveillance, Epidemiology, and End Results (SEER) database and enrolled female primary non-metastatic cases. Patients were subdivided into seven groups, and analyses of the associations between age and overall survival (OS) and breast cancer-specific survival (BCSS) were carried out using the Kaplan-Meier method and Cox regression model, respectively. We also assessed differences in survival among three specific age groups, using the ages of 30 and 50 years as cut-offs. Stratified analyses regarding race, histology, grade, stage and hormone receptor status were also carried out. Results A total of 133,057 female patients diagnosed with breast cancer from 2004 to 2008 were included in the current study (6.4% <40 years), Women aged 40 to 49 years and 60 to 69 years exhibited significantly better OS and BCSS, respectively (log-rank, p<0.001), than their counterparts in other groups. Middle-aged women exhibited distinctly better OS (log-rank, p<0.001) and BCSS (log-rank, p<0.001) than their counterparts in the other two age groups. Following adjustments for potential confounding factors, middle-age at breast cancer diagnosis was shown to be an independent predictor of favourable outcomes in terms of OS, but not BCSS (for OS, HR, 0.92; 95%CI, 0.87–0.98; p = 0.007; for BCSS, HR, 0.94; 95%CI, 0.80–1.01; p = 0.075, using the young group as reference). Stratified analysis showed that middle-age was significantly associated with increased survival, except among patients with stage III disease, and that elderly women faced worse prognoses than younger patients. Conclusion Our results indicate that younger breast cancer patients exhibit more aggressive disease than older patients. Middle-aged patients exhibit better OS and BCSS than young and elderly patients but exhibit BCSS rates similar to those of young patients after adjustments for confounders. Stratified analysis demonstrated that middle-aged patients exhibited better survival than young patients, with the exception of patients with stage III disease. An age of 60 years or more was a significant independent predictor of a poor prognosis.

MiR-944 functions as a novel oncogene and regulates the chemoresistance in breast cancer.

MircroRNAs are emerging as critical regulators in carcinogenesis and chemoresistance in multiple cancer types. In this study, we observed that the miR-944 level was upregulated in breast cancer patients’ serum and tumor tissues, suggesting that miR-944 is a tumor promoter in breast cancer. To investigate the role of miR-944, we performed gain- and loss-of-function experiments in vitro. We then demonstrated that miR-944 promotes cell proliferation and tumor metastasis in breast cancer cell lines. Furthermore, we indicated that miR-944 is associated with cisplatin resistance by targeting BNIP3. Knockdown of the miR-944 by specific inhibitors significantly increased the cytotoxicity of cisplatin in cisplatin-resistant MCF-7 cells (MCF-7/R). Importantly, we found that the sensitization of miR-944 inhibitors to cisplatin cytotoxicity was abolished by BNIP3 siRNA which decreased the expression of BNIP3 gene. Finally, we demonstrated that miR-944 inhibitors promoted the loss of mitochondrial membrane potential (MMP) caused by cisplatin in MCF-7/R cells, resulting in the release of mitochondria-derived apoptogenic proteins into cytoplasm, and then, the caspase-3 was activated. In summary, our study showed that miR-944 functions as a novel oncogene and regulates the cisplatin resistance in breast cancer. The miR-944-BNIP3-MMP-caspase-3 pathway might be a novel target for the chemotherapy of breast cancer.

Honokiol synergizes chemotherapy drugs in multidrug resistant breast cancer cells via enhanced apoptosis and additional programmed necrotic death

Multidrug resistance (MDR) is a major challenge in cancer therapy. Apoptosis tolerance is one of the key mechanisms of MDR. Honokiol, a small-molecule pharmacologically active component, exhibits competent cytotoxicity in a variety of human cancer cells through apoptosis and other forms of programmed cell death (such as programmed necrosis). Although much work has been done on its antitumor effects, little attention has been paid on systemic evaluation of efficacy of honokiol combined with other chemotherapeutic agents, especially in drug‑resistant cell lines. Here, we systematically and quantitatively assess its combinational effect with different chemotherapeutic agents using the combination index (CI) equation. We found that honokiol synergized with chemotherapeutic agents both in sensitive and resistant, solid and non-solid (MCF-7, HL-60, MCF-7/ADR and HL-60/ADR) cell lines. Honokiol (40 µg/ml) induced necrotic cell death in MCF-7/ADR cells with characterized morphological and biochemical features. Co-incubation with honokiol and etoposide (VP-16) activated a complex death modality, which was composed of necrotic cell death and apoptosis. This dual-death pathway was shut down when pretreated with pan-caspase inhibitor (z-VAD-fmk) and cyclophilin D inhibitor (cyclosporin A). Western blot analysis results proved that honokiol also enhanced VP-16-induced apoptosis potentially via blocking nuclear factor‑κB (NF-κB) activation. Our data for the first time quantitatively demonstrate that honokiol synergizes frequently-used chemotherapeutic agents via enhanced apoptosis and additional programmed necrotic death. These findings indicate a promising way to circumvent MDR and apoptosis tolerance.

Diaphragmatic perforation with colonic herniation due to hepatic radiofrequency ablation: A case report and review of the literature

Radiofrequency ablation (RFA) has been widely accepted as an alternative treatment for unresectable primary and metastatic hepatic tumors, with satisfactory rates of local response and significant improvements in rates of overall survival. Numerous large series studies have shown that RFA is safe and effective, with a low mortality rate and a low major complication rate. Major complications, including diaphragmatic perforation and hernia, have rarely been previously reported. The current case report presents a case of diaphragmatic hernia with perforation of the incarcerated colon in the thoracic cavity 12 months following hepatic RFA, and reviews nine previously reported cases of diaphragmatic hernia. Comprehensive analysis of the nine cases demonstrated possibilities leading to diaphragmatic hernia following diaphragmatic thermal injury as a consequence of hepatic RFA. Clinicians and radiologists must consider diaphragmatic thermal damage following hepatic RFA for liver tumors adjacent to the diaphragm, particularly for patients with symptoms of ileus, dyspnea, chest pain, pleural effusion and right shoulder pain.

New clinical application of high-intensity focused ultrasound: local control of synovial sarcoma

High-intensity focused ultrasound (HIFU) is playing an increasingly important role in cancer therapy. Primary synovial sarcomas of the chest wall are extremely rare. We report the first case of noninvasive HIFU therapy for the control of synovial sarcoma. A 51-year-old man was diagnosed with spindle cell sarcoma on the left chest wall through lumpectomy. After four cycles of chemotherapy, local recurrence of the sarcoma was detected. Subsequent extended resection confirmed synovial sarcoma. After five cycles of a new chemotherapy option, the sarcoma relapsed again. Then the patient received five courses of HIFU; this completely ablated the sarcoma without complications. No chemotherapy, radiotherapy, or biological therapy has been applied since. Now the patient is stable and has a high quality of life.

Enhanced lymph node retrieval from colorectal cancer resections using a simple lymphatic staining method.

BACKGROUND/AIMS This study evaluated the effect of lymphatic staining on the number of lymph nodes (LNs) examined and staging in patients with colorectal cancer. METHODOLOGY Sixty-two consecutive specimens from patients with colorectal cancer resected between February 2009 and April 2010 were randomized to the stained group or the control unstained. Differences in the retrieval, number and size of nodes, and time for retrieval were measured. RESULTS LN harvest differed significantly with 30±12 and 13±5 (p<0.01) nodes in the stained and the control groups, respectively. Insufficient LN harvest (less than 12 nodes) occurred in 14 cases of the control group and only in 1 case of the stained group (p<0.01). Metastases were confirmed in 57 LNs occurring in 17 cases of the stained group and in 39 nodes occurring in 15 cases of the control group. The mean time for LN retrieval in the stained and control groups were comparable, 27.6±6.9min and 24.7±6.0min (p>0.05), respectively, yet there was a significant difference in the number of LNs (<2mm) (294 vs. 59, respectively, p<0.01) as well as in the number of LNs 2-5mm in size (474 vs. 220, respectively, p<0.01). CONCLUSIONS By lymphatic staining method, more and smaller LNs could be detected, which significantly improved the LN harvest of resected colorectal specimens and reduced cases of insufficient LN harvest.

Cyclophilin D modulates cell death transition from early apoptosis to programmed necrosis induced by honokiol

Honokiol is a pharmacologically active small molecule with multifunctional antitumor effects. Although plenty of literature is available on honokiol-triggered apoptosis and programmed necrosis, few studies have investigated the potential existence of death mode transition from apoptosis to programmed necrosis. In the current study, we demonstrated that the necrotic cell population (PI-positive) gradually increased and the early-stage apoptotic cell population (PI-negative and AV-positive) decreased in a dose- and time-dependent manner following honokiol treatment. Furthermore, we demonstrated that these PI-positive cells were under necrotic cell death, since no late-apoptosis characteristics including conspicuous chromatin condensation or DNA ladder patterns were detected. These results demonstrated that cells suffered death mode transition from early-stage apoptosis to programmed necrosis with the increase of honokiol dose or treatment time. The protein expression of RIP3 markedly increased in parallel with HNK-triggered death mode transition, while the expression of RIP1 decreased. Cyclophilin D expression increased during cell death mode transition, and inhibition of cyclophilin D by cyclosporin A clearly blocked HNK-triggered programmed necrosis. These data indicated that honokiol-induced programmed necrosis and death mode transition are potentially RIP3‑dependent, cyclophilin D-regulated. Further results showed that blocked cyclophilin D by cyclosporin A inhibited HNK-induced necrosis, but did not affect HNK-induced RIP3 overexpression. This indicated that cyclophilin D was a potential modulator at downstream of RIP3. In conclusion, honokiol triggers a potential RIP3-dependent cell death mode transition from early-stage apoptosis to programmed necrosis, which is highly regulated by cyclophilin D.